Cloned (Comment) | Organism |
---|---|
overexpression in Escherichia coli strain RB791, amino acid determination, incorrect positioning of disulfide bonds, the recombinant enzyme is less thermostable and thermophilic than the native enzyme | Saccharolobus solfataricus |
Crystallization (Comment) | Organism |
---|---|
from recombinant enzyme, hanging drop-vapour diffusion method, protein solution, 7-10 mg/ml, 18°C, reservoir solution for native crystals: Tris-HCl 10 mM, pH 7.4, 28-30% dioxane, 12% 2-methyl-2,4-pentanediol, 0.12 M MgCl2, 0,04 M NaCl, for crystals of enzyme complexed with substrates or sulfate and phosphate ions, substrates are added and NaCl is exchanged for MgSO4 or NH4Cl and KH2PO4, respectively, X-ray structure determination and analysis | Saccharolobus solfataricus |
General Stability | Organism |
---|---|
phosphate, and less efficiently also arsenate and sulfate, stabilize the recombinant enzyme against inactiviation by temperature, SDS, urea, and proteolytic enzymes | Saccharolobus solfataricus |
recombinant enzyme, 90°C in 2% SDS, 30 min, loss of 60% activity | Saccharolobus solfataricus |
recombinant enzyme, 90°C in 8 M urea, 30 min, loss of 70% activity | Saccharolobus solfataricus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
proteinase K | recombinant enzyme, 10% remaining activity after 4 h at 37°C, phosphate protects | Saccharolobus solfataricus | |
Subtilisin | recombinant enzyme, 24% remaining activity after 4 h at 37°C, phosphate protects | Saccharolobus solfataricus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
27000 | - |
6 * 27000, recombinant enzyme, crystal structure analysis | Saccharolobus solfataricus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharolobus solfataricus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant from Escherichia coli | Saccharolobus solfataricus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5'-methylthioadenosine + phosphate | binding of phosphate and 5-methylthioribose 1-phosphate to the enzyme induces a conformational transition that stabilizes the folded structure of the enzyme | Saccharolobus solfataricus | adenine + 5-methylthio-D-ribose 1-phosphate | - |
? | |
5'-methylthioadenosine + phosphate | 5'-methylthioadenosine and adenine form ternary complexes with the enzyme only in presence of phosphate and methylthioribose 1-phosphate, respectively | Saccharolobus solfataricus | adenine + 5-methylthio-D-ribose 1-phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | 6 * 27000, recombinant enzyme, crystal structure analysis | Saccharolobus solfataricus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
100 | - |
recombinant enzyme, 1 h, 95% remainig activity in presence of 100 mM phosphate | Saccharolobus solfataricus |
110 | - |
recombinant enzyme, 10 min, 50% remaining activity in absence and 90% remaining activity in presence of 100 mM phosphate | Saccharolobus solfataricus |
120 | - |
recombinant enzyme, 10 min, no activity in absence and 50% remaining activity in presence of 100 mM phosphate | Saccharolobus solfataricus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Saccharolobus solfataricus |